BACKGROUNDCurrently anti-inflammatory therapy with steroids for allergic rhinitis need long-term repeated administration, although it is effective. Gene therapy is being suggested to substitute it. The aim of this study was to investigate nonviral vector mediated exogenous gene expression in COS-7 cells in vitro and the effect of intranasal mouse interleukin (mIL)-12 transgene expression on allergen induced eosinophil infiltration of nasal mucosa in a murine model of allergic rhinitis.

METHODSIn vitro COS-7 cells were infected with Epstein-Barr virus (EBV)/lipoplex. The expression of IL-12 p70 in cell culture supernatant was examined by enzyme-linked immunosorbent assay (ELISA). In mice with ovalbumin (OVA) induced allergic rhinitis, EBV/lipoplex was administered by nasal drops before OVA challenge once a day from day 1 to day 10. The expression of IL-12 mRNA and protein, the change of eosinophil count in nasal mucosa and serum total IgE were measured 24 hours after the last challenge.

RESULTSEBV/lipoplex could effectively transfect COS-7 cells. The expression of IL-12 p70 in cell culture supernatant was significantly more than in blank control. IL-12 via EBV plasmid vector transduction could be overexpressed in vivo. In pGEG.mIL-12 treated models, the nasal mucosa revealed a high level of widespread mIL-12 transduction by immunohistochemistry and in situ hybridization. Histological evaluation revealed marked suppression of eosinophil infiltration in nasal mucosa. The eosinophil count in allergic rhinitis group [(26.5 +/- 9.8)/high-power field (HPF)] was significantly increased over control group [(0.40 +/- 0.52)/HPF] (F = 56.94, P < 0.01), while the count in IL-12 gene therapy group [(4.60 +/- 2.63)/HPF] was significantly less than that of allergic group (F = 56.9, P < 0.01). Serum total IgE between in gene therapy mice [(88.83 +/- 6.71) ng/ml] and allergic rhinitis mice [(103.1 +/- 5.7) ng/ml] showed a significant difference (F = 1216, P < 0.05).

CONCLUSIONSNonviral EBV plasmid vector, pGEG.mIL-12 was able to overexpress exogenous gene both in vitro and in murine nasal mucosa in vivo. IL-12 overexpression via EBV/lipoplex could stem allergen induced eosinophil infiltration in nasal mucosa in murine models of allergic rhinitis, which may suggest a new cytokine immunogenetic therapy for allergic rhinitis.

Administration, Intranasal ; Animals ; COS Cells ; Cercopithecus aethiops ; Eosinophilia ; therapy ; Genetic Therapy ; Herpesvirus 4, Human ; genetics ; Immunoglobulin E ; blood ; Interleukin-12 ; genetics ; Lipids ; administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy ; Rhinitis, Allergic, Seasonal ; therapy

OBJECTIVETo observe the effects of gene therapy and epigenetic therapy on the tumor growth of laryngeal carcinoma and the underlying mechanisms.

METHODSThe animal model of human laryngeal carcinoma was established by the subcutaneous inoculation of Hep-2 cells at the right armpit of BALB/c nu/nu mice. The tumor-bearing mice were randomized into 4 groups, p53 therapy group(rAd-p53), epigenetic therapy group(5-aza-dC), combination therapy group (rAd-p53+5-aza-dC) and control group. The gene and protein expressions of molecular markers p53 and E-cadherin were detected by FQ-PCR and immunohistochemistry.

RESULTSBy the day 20 of the treatments, the mean tumor volumes were(106.09 ± 24.40)mm(3) in p53 therapy group, (166.55 ± 40.11) mm(3) in epigenetic therapy group, (126.11 ± 22.49) mm(3) in combination therapy group,and (252.83 ± 54.09) mm(3) in control group. Both gene therapy (F = 37.30, P < 0.05) and epigenetic therapy (F = 4.79, P < 0.05) inhibited the growth of xenografted tumors, with an interaction effect (F = 22.01, P < 0.05) between the two groups. The integral optical density value of p53 protein expression of p53 therapy group (628.07 ± 95.16) was significantly higher than that of combination therapy group (494.76 ± 100.22), (t = 8.72, P < 0.05). The integral optical density values of E-cadherin protein expression were 558.89 ± 97.58 in p53 therapy group, 380.41 ± 90.60 in epigenetic therapy group, 494.76 ± 102.88 in combination therapy group,and 162.60 ± 40.38 in control group respectively, indicating the enhancements of E-cadherin protein expression by gene therapy (F = 45.24, P < 0.05) or epigenetic therapy(F = 5.73, P < 0.05)and the existence of interaction effect (F = 21.82, P < 0.05) between gene therapy and epigenetic therapy. The expression levels of p53 gene were 4.43 ± 0.12 in p53 therapy group, 1.06 ± 0.11 in epigenetic therapy group, 3.51 ± 0.10 in combination therapy group,and 1.09 ± 0.11 in control group, respectively, showing an interaction effect between gene therapy and epigenetic therapy (F = 298.11, P < 0.05). The expression levels of E-cadherin gene were 4.50 ± 0.34 in p53 therapy group, 2.02 ± 0.16 in epigenetic therapy group, 2.99 ± 0.12 in combination therapy group, and 1.00 ± 0.11 in control group, respectively. The expression of E-cadherin gene was enhanced by gene therapy (F = 329.12, P < 0.05)or epigenetic therapy(F = 88.57, P < 0.05), with an interaction effect between the two therapies (F = 122.17, P < 0.05).

CONCLUSIONSXenografted tumors of human laryngeal carcinoma cells are inhibited by gene therapy, the epigenetic therapy and the combination therapy. The gene therapy was significantly better than the epigenetic therapy or the combination therapy. There might be antagonistic effect between p53 and 5-aza-dC.

Animals ; Cadherins ; metabolism ; Carcinoma ; therapy ; Cell Line, Tumor ; Combined Modality Therapy ; Epigenomics ; Genetic Therapy ; Humans ; Laryngeal Neoplasms ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Tumor Suppressor Protein p53 ; metabolism

BACKGROUNDGene therapy and epigenetic therapy have gained more attention in cancer treatment. However, the effect of a combined treatment of gene therapy and epigenetic therapy on head and neck squamous cell carcinoma have not been studied yet. To study the mechanism and clinical application, human laryngeal carcinoma cell (Hep-2) tumor-bearing mice were used.

METHODSA xenograft tumor model was established by the subcutaneous inoculation of Hep-2 cells in the right armpit of BALB/c nu/nu mice. The mice with well-formed tumor were randomly divided into six groups. Multisite injections of rAd-p53 and/or 5-aza-dC were used to treat tumor. Tumor growth was monitored by measuring tumor volume and growth rate. p53 and E-cadherin protein levels in tumor tissues were detected by immunohistochemical staining. The mRNA levels were monitored with FQ-PCR.

RESULTSGene therapy was much more effective than single epigenetic therapy and combined therapy. The gene therapy group has the lowest tumor growth rate and the highest expression levels of p53 and E-cadherin.

CONCLUSIONSThe combined treatment of gene and epigenetic therapy is not suggested for treating head and neck carcinoma, because gene therapy shows an antagonistic effect to epigenetic therapy. However, the mechanisms of action are still unclear.

Animals ; Azacitidine ; analogs & derivatives ; therapeutic use ; Cadherins ; analysis ; DNA Modification Methylases ; antagonists & inhibitors ; Epigenesis, Genetic ; Genes, p53 ; Genetic Therapy ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Tumor Suppressor Protein p53 ; analysis ; Xenograft Model Antitumor Assays

OBJECTIVETo analyze the correlation between airborne pollen concentrations and symptoms in patients with pollen allergic rhinitis.

METHODSDurhum sampler was used to collect the pollen concentration and species from June to September in 2011. The clinical skin prick test (SPT) data were analyzed. The patients with pollen allergic rhinitis were divided into pure pollen allergic rhinitis group (pollen group) and pollen combined perennial allergens allergic rhinitis group (combined group). Symptom scores of patients were assessed, and correlation between pollen concentration and onset of symptoms of patients were analyzed. SPSS 16.0 software was used to analyse the data.

RESULTSWhile the peak of Summer-Autumn pollen concentration appeared from August 20 to September 15, the major pollen included Artemisia L, Chenopodium album and Humulus scandens. The peak of pollen concentration in one day reached 638/1000 mm(2). The patients taken SPT from June to September accounted for 51.9% of the patients in whole year, among which SPT pollen positive patients were 1509, 60.7% of all SPT positive patients. The amount and rate of SPT positive patients showed significant correlation with pollen concentration(r value were 0.90 and 0.99, both P < 0.05). Onset of symptoms in two groups was correlated with pollen concentration in Summer-Autumn. Symptoms of cough in combined group showed more severe compared with patients with pollen group (t = 2.36, P < 0.05).

CONCLUSIONSPollen concentration has a major effect on onset of symptoms of allergic rhinitis. Airborne pollen monitoring has important preventive and therapeutic significance on patients with allergic rhinitis.

Adolescent ; Adult ; Air ; analysis ; Allergens ; immunology ; Child ; Environmental Monitoring ; Humans ; Middle Aged ; Pollen ; immunology ; Rhinitis, Allergic, Seasonal ; diagnosis ; epidemiology ; Seasons ; Skin Tests ; Young Adult

OBJECTIVETo observe the histopathological finding of bone remodeling in rabbit sinusitis model at different time and the tendency, and to discuss the effect of bone in the pathogenesis of sinusitis.

METHODSFirst, the rabbit sinusitis model was made, then the experimental animals were divided into 3 groups according to the time of infection. There were 8 rabbits in each experimental group, and 4 rabbits in the control group. The sinus specimen were collected, embedded and stained. The bone in the inoculating side and noninoculating side was scored, and the bone in inoculating side was evaluated quantitatively and semiquantitatively. The parameters included the thickness of mucosa, mucoperiosteum, the density of osteoblast, the amount of osteoclast.

RESULTSThe average bone score in the inoculating side was 2.250, 2.875, 2.875; in the noninoculating side was 1.625, 2.250, 2.500. Between group A and the control group, the difference of all three parameters had statistical significance. Between group B and group A, the difference of the thickness of mucosa and the density of osteoblast had statistical significance. Between group C and group B, none of the three parameters had statistical significance.

CONCLUSIONSBacterial sinusitis can lead to bone remodeling, obvious bone destroy can occur at the early phase, then the bone proliferation follows. These results demonstrate that bone remodeling is one of the basic histopathological characters of CRS and might be the reason to lead CRS to a constant and chronic process of inflammation.

Animals ; Bone Remodeling ; Disease Models, Animal ; Inflammation ; Nasal Bone ; pathology ; Rabbits ; Sinusitis ; pathology

OBJECTIVETo investigate the histopathological changes in cricoarytenoid joints in 32 animal models. The characteristic histopathological changes of arytenoid cartilages after recurrent nerve paralysis were evaluated.

METHODSSixteen dogs (32 vocal folds, 8 as normal control) were divided into different animal models of recurrent nerve paralysis as transection, half-section, ligation, or crush. The histopathological finds of arytenoid cartilages were analysed.

RESULTSArytenoid cartilages showed fibrin (12/24), disruption of the fibrous membrane (9/24), fibrillation (7/24) and degenerative changes in their joint surface structure (3/24) at various levels of intensity. The fibrin and disruption of the fibrous membrane were found 1 month after injury, and all changes appeared in 6 months. The fibrillation and arytenoid cartilages degenerative changes revealed in transaction group and ligation group, and became stronger in time of 6 months. The correlation among the fibrillation ratio and the normal control was positive (t were 6.23 and 3.65, P < 0.01). The correlation among the number of cellular of arytenoid cartilages and the normal control was positive (t = 2.78, P < 0.05). The fibrillation (7) and arytenoid cartilages degenerative changes (3) revealed in vocal fold fixation to influence the recovery of laryngeal function.

CONCLUSIONSThe histopathological change of cricoarytenoid joint after recurrent nerve paralysis was related to the severity of neural injury. Influence the recovery of laryngeal function more often from 6 months.

Animals ; Arytenoid Cartilage ; pathology ; Cricoid Cartilage ; pathology ; Disease Models, Animal ; Dogs ; Joints ; pathology ; Recurrent Laryngeal Nerve ; pathology ; Recurrent Laryngeal Nerve Injuries

OBJECTIVETo evaluate the histopathologic changes of the ethmoid bone in CT scan typing of chronic rhinosinusitis (CRS), and investigate relations with outcomes of endoscopic surgery.

METHODSOne hundred and twelve random CRS patients undergoing endoscopic sinus surgery at Tongren hospital were prospectively evaluated. According to the preoperative CT scan, these patients were divided into three groups of CT scan typing. The bony changes and bone remodeling of ethmoid bone were graded and relations with outcomes of endoscopic surgery were investigated.

RESULTSThe CT scan typing results of ethmoid sinus of CRS were as followed: 22.3% (type III), 39.3% (type II), and 38.4% (type I). Comparison of bone remodeling and postoperative endoscopic scores between the three groups revealed a significant difference (P = 0.01, P = 0.02). A trend toward more advanced bone remodeling grade in association with a higher postoperative endoscopic scores was identified, with a statistically significant correlation.

CONCLUSIONThe Chinese CT scan typing of CRS has its pathological basis and can a useful method to predict the surgical outcomes of CRS. The new bone formation or bony tissue remodeling of sinus often represents pathogenesis of CRS and implicates the prognosis of sinus surgery.

Adult ; Chronic Disease ; Ethmoid Sinus ; diagnostic imaging ; pathology ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Prospective Studies ; Sinusitis ; classification ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo evaluate the effect and safety of the formulation and dosage of aluminium adjuvant, Al(OH)(3), in the preparation of allergic rhinitis animal model.

METHODSSixty health BALB/c mice were divided randomly into 6 groups. Al(OH)(3) powder (5 mg) was used in one group, Al(OH)(3) colloid gel of different concentration (0.5 - 5 mg) was used in four groups, and normal saline was used in the control group. Ovalbumin injection and nasal topical challenge were used in the 5 testing groups to induce allergic rhinitis in mice. Normal saline was used in the control group.

RESULTSTypical allergic rhinitis symptoms including frequent nasal scratching, and edema of peri-nasal mucosa were found in mice of the 5 mg Al(OH)(3) powder group. Eosinophils accumulation, goblet cells hyperplasia and hypersecretion were found in the mucosa of lateral nasal wall and inferior nasal turbinate. Neither obvious allergic rhinitis symptom, nor eosinophils accumulation in nasal mucosa was observed in the Al(OH)(3) colloid gel groups. Hemorrhagic ascites and lots of white nodules (foreign body granuloma) formation were found in the liver, spleen, and kidney of all mice of the 5 mg Al(OH)(3) colloid gel group. Five out of 10 mice of the 2 mg Al(OH)(3) colloid gel group exhibited above signs but of lower grade. Despite dispersed fine white sediment in the liver and mesentery, no obvious ascites was found in mice of the 1 mg and 0.5 mg Al(OH)(3) colloid gel groups.

CONCLUSIONSAl(OH)(3) powder, 5 mg, is effective and safe in the preparation of allergic rhinitis animal model. Al(OH)(3) colloid gel of different concentration (0.5 - 5 mg) may cause side effects such as foreign body granuloma.

Adjuvants, Immunologic ; administration & dosage ; adverse effects ; Administration, Intranasal ; Aluminum Hydroxide ; administration & dosage ; adverse effects ; Animals ; Disease Models, Animal ; Female ; Mice ; Mice, Inbred BALB C ; Rhinitis, Allergic, Perennial ; Rhinitis, Allergic, Seasonal

OBJECTIVETo analyze the impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons (ORN), and discuss the reliability of this experimental model.

METHODSAfter olfactory nerve transection of mice, anterograde horseradish peroxidase (HRP) tracing was performed to confirm the completion of nerve transection. On 8 h, 2 d, 3 d and 5 d after surgery, TdT mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) was used to observe the apoptosis of ORN, while relative semi-quantitative RT-PCR and immunohistochemistry were used to reflect the expression of olfactory marker protein (OMP, special marker of mature ORN) in olfactory epithelium.

RESULTSNo HRP label was observed in olfactory bulb after olfactory nerve transaction. Both TUNEL-positive and OMP-positive cells were ORN. After the surgery, TUNEL-positive cells increased remarkably and peaked on 2 d after the surgery. Meanwhile OMP mRNA in olfactory epithelium began to decrease markedly till 5 d after the surgery, and the olfactory epithelium got thinner accordingly.

CONCLUSIONSThis experimental model can be used reliably to sever mice olfactory nerve and manipulate simultaneous apoptosis of mice ORN.

Animals ; Apoptosis ; Denervation ; Mice ; Mice, Inbred C57BL ; Olfactory Nerve ; cytology ; metabolism ; surgery ; Olfactory Receptor Neurons ; metabolism ; pathology

OBJECTIVETo investigate whether the local application of IL-12 gene with EBV-plasmid vector to nasal mucosa could prevent allergic inflammation in murine allergic rhinitis model.

METHODSThirty-six BALB/C mice were randomly divided into allergic rhinitis group gene therapy group and control group. In mice with OVA-induced allergic rhinitis, the EBV/lipoplex (a novel cationic lipid combined with EBV-plasmid vector, pGEG. mIL-12) was locally administered into nasal mucosa before OVA challenge. The expression of IL-12 mRNA and protein, the change of eosinophilia and mast cell, and Th2 cytokine production in the nasal mucosa were measured.

RESULTSThe amounts of IL-12 mRNA positive cells and IL-12 positive cells in nasal mucosa of gene therapy group were significantly higher than that of allergic rhinitis group (P < 0.01 and P < 0.05). The amount of eosinophils, mast cells, and the level of IL-5 expression in nasal mucosa in allergic rhinitis group were significantly higher than those in gene therapy group and control group (P < 0.01). The level of total IgE of peripheral blood in allergic rhinitis group was significantly higher than that in gene therapy group and control group (F = 1216.21, P < 0.01).

CONCLUSIONSThese findings indicated that IL-12 mRNA and protein were expressed effectively after the local administration of pGEG. mIL-12 in the nasal mucosa. The local application of pGEG. mIL-12 is effective in modulating nasal allergic response and may be a convenient method for future approach to allergic rhinitis.

Animals ; Genetic Therapy ; Genetic Vectors ; Interleukin-12 ; genetics ; therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy